Flow Cytometry Lab
The Flow Cytometry laboratory is housed in Mosier Hall, Room K208 of the College of Veterinary Medicine Complex. This multi-user core laboratory is equipped with state of the art flow cytometry analyzers by Sony (EC800) and Becton Dickinson (FACSCalibur), a multiplex MagPix instrument, and fluorescent activated cell sorting equipment by Beckman Coulter (MoFLO XDP).
Hours of Operation: M – F 8 – 5 pm, Call to schedule a time on the instruments
Usage Fees:
Bench top analyzers $20.00/15 minutes for acquisition
Assisted acquisition and analysis $30/15 minutes by staff
Sorting $100/hour.
The MoFlo XDP is operated solely by a member of the staff at the facility, after consultation with the client.
Training: $100 for 2 people (2 hr duration)
Comprehensive training on the FACSCalibur or EC800 will be offered to users on request including instrument orientation, sample acquisition, data analysis, experiment setup, and instrument troubleshooting
E-mail or call for consultation:
Director: |
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Melinda J. Wilkerson, DVM PhD |
Ph: 785-532-4818 |
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Research Associate and Dedicated Cell Sorter Operator: |
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Catherine Ewen, PhD |
Ph: 785-532-4617 |
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FAX Number: 785-532-4072 |
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Research Assistant (Flow Cytometry certified operator): |
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Anushka George, MS |
Ph: 785-532-4617 |
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FAX Number: 785-532-4072 |
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Sony EC800:
The Sony EC800 is equipped with four lasers (405, 488, 561, and 640) with 6-color capabilities. This instrument can determine size, complexity, and electronic volume. This flow cytometer can sample directly from a variety of plate or tube formats (ependorf tubes, falcon tubes, culture well plates, and tube rack) and is fully automated. The 405 laser allows the flexibility to use violet dyes. The 561 lasers provide the capability to excite a variety of fluorescent proteins (mCherry, mKate, link to fluorescent pdf chart).
FACSCalibur:
The FACSCalibur is equipped with two lasers that emit light at 488 nm and 630 nm. There are three detectors (FL1, FL2, FL3) for the 488 laser and one detector (FL4) for the 630 laser. This two laser cytometer allows the operator to analyze up to 4-color and 2 size parameters of cells or fluorescent-labeled particles. FACSCalibur is interfaced with a Macintosh computer and a publication quality color printer. Cell Quest and ModFit analysis software is available on two computers to analyze flow cytometry data including DNA and proliferation assays.
MoFLO XDP:
The MoFLO XDP is a “state of the art” analytical flow cytometer with high speed sorting capabilities. The MoFlo has three lasers (argon/UV/red diode) with the capability to perform 6-color and 2 size parameter analyses and to routinely sort cells or particles at speeds up to 70,000 cells/second.
The research capabilities of this facility are very diverse and only limited by the imagination, available fluorochromes, and capabilities of the instruments. Examples of the research experiments that can be performed at this facility include but are not limited to: apoptosis studies, immunophenotyping, DNA content and viability of spermatozoa and other cell types, intracellular calcium levels, respiratory burst and phagocytic function, cell cycle, enzymatic activity, sorting of individual subpopulations of cells from a mixed population, sorting of GFP expressing cells from mixed cultures, and analysis of PCR products attached to microspheres. (Link to ppt slides for basic theory and applications)
MagPix:
The MagPix by Millipore can analyze up to 50 analytes in a single sample. Examples are cytokine, chemokine, and serological panels. Call for consultation and quote for usage.
http://www.millipore.com/bmia/flx4/bmia_instruments#tab1=2
Additional educational resource (Tutorials on flow cytometry)
http://probes.invitrogen.com/resources/education/tutorials/4Intro_Flow/player.html